Anti-Merlin (NF2) Antibody, clone 1C4

Merlin (UniProt P35240; also known as Moesin-ezrin-radixin-like protein, Neurofibromin-2, Schwannomerlin, Schwannomin) is encoded by the NF2 (also known as SCH) gene (Gene ID 4771) in human. Originally identified as a tumor suppressor gene in the familial Neurofibromatosis type 2 (NF2) cancer patients, NF2 encodes the FERM (Ezrin, Radixin, Moesin) domain-containing merlin that plays a negative regulating role against cell growth and proliferation by functioning as an upstream activator of the Hippo pathway. Merlin directly activates Mst1/2 and recruits Lats1/2 to the plasma membrane for phosphorylation by Mst1/2. The phosphorylated Lats1/2 in turn inhibits YAP/TAZ transcription activity by phosphorylating YAP/TAZ and causes its cytoplasmic sequestration. Merlin also translocates to the nucleus, where it inhibits E3 ubiquitin ligase CRL4(DCAF1), thereby preventing it from ubiquitinating and downregulating Lats1/2. In its inactive state, the C-terminal tail domain (CTD; a.a. 507-595) of merlin is bound intramolecularly to its FERM domain (a.a. 19-313). Angiomotin (AMOT) induces merlin activation by binding to an elongated helical fragment within merlin CTD and subsequently releases the auto-inhibited conformation of Merlin. Merlin CTD can also undergo phosphorylation on Ser518 by p21-activated kinase (PAK) or cyclic AMP-dependent protein kinase (PKA), resulting in a weakened AMOT binding and a stabilization of merlin in its auto-inhibited state.

Clone 1C4 detected merlin (NF2), but not any of the three ERM proteins, ezrin, radixin, and moesin (Gonzalez-Agosti, C., et al. (1996). Oncogene. 13(6):1239-1247). Clone 1C4 targeted epitope is present in human merlin (NF2) spliced isoforms 1 to 6, but not 7, 9, or 10 as reported by UniProt (P35240). Reactivity toward human spliced isoform 8 is possible, but has not been confirmed.

Recombinant protein corresponding to the C-terminal half of human merlin (NF2) isoform II (UniProt spliced isoform 3; P35240-3).

Anti-Merlin (NF2), clone 1C4, Cat. No. MABN1786, is a highly specific mouse monoclonal antibody that targets Merlin (NF2) and has been tested in Immunocytochemistry, Immunoprecipitation, and Western Blotting.

Immunocytochemistry Analysis: A representative lot immunostained % paraformaldehyde-fixed, 0.2% Triton X-100-permeabilized wild-type, but not Nf2-knockout, mouse liver-derived epithelial cells (LDCs) by fluorescent immunocytochemistry. EGF-induced EGFR internalization was observed among Nf2-/-, but not Nf2+/+, LDCs (Curto, M., et al. (2007). J. Cell Biol. 177(5):893-903).

Immunocytochemistry Analysis: A representative lot detected merlin cellular distribution and HEI10 co-localization in a cell cycle-dependent manner by fluorescent immunocytochemistry staining of 3.5% paraformaldehyde-fixed U2OS human osteosarcoma cells (Grönholm, M., et al. (2006). Oncogene. 25(32):4389-4398).

Immunocytochemistry Analysis: A representative lot detected merlin cellular localization by fluorescent immunocytochemistry staining of 3.5% paraformaldehyde-fixed embryonic E16 rat neurons. Merlin was seen co-localized with RI to the cell body and extensions in a punctate pattern (Grönholm, M., et al. (2003). J. Biol. Chem. 278(42):41167-41172).

Immunocytochemistry Analysis: A representative lot detected merlin cellular localization by fluorescent immunocytochemistry staining of 4% paraformaldehyde-fixed, 0.1 NP-40-permeabilized human fibroblasts and primary meningioma cells. Merlin co-localized with F-actin at the leading and ruffling edges, but not at the stress fiber. No merlin co-localization with ezrin or moesin was observed (Gonzalez-Agosti, C., et al. (1996). Oncogene. 13(6):1239-1247).

Immunoprecipitation Analysis: A representative lot co-immunoprecipitated N-WASP with merlin from HEK293T cell lysate (Manchanda, N., et al. (2005). J. Biol. Chem. 280(13):12517-12522).

Western Blotting Analysis: A representative lot detected the presence of wild-type, but not L64P, merlin in the EGFR immunoprecipitates from Nf2-/- MEFs infected by adenovirus to express either wild-type or L64P merlin (Curto, M., et al. (2007). J. Cell Biol. 177(5):893-903).

Western Blotting Analysis: A representative lot detected a reduced merlin level in EGFR immunoprecipitate from mouse liver-derived epithelial cells (LDCs) upon shRNA-mediated NHE-RF1, but not NHE-RF2 knockdown. Merlin association with Ezrin was not affected by NHE-RF1 knockdown (Curto, M., et al. (2007). J. Cell Biol. 177(5):893-903).

Western Blotting Analysis: A representative lot detected a ~72 kDa merlin band in rat newborn fibroblast (RNF) and S-16 rat Schwann cell lysates, as well as a ~66 kDa merlin band in murine NIH/3T3 and human MRC-5 fetal fibroblast lystes without cross-reactivity toward the three ERM protein family members, ezrin, radixin, and moesin (Gonzalez-Agosti, C., et al. (1996). Oncogene. 13(6):1239-1247).

Research Category
Neuroscience

Evaluated by Western Blotting in HEK293 cell lysate.

Western Blotting Analysis: 4 µg/mL of this antibody detected Merlin (NF2) in 10 µg of HEK293 cell lysate.

~70 kDa observed. 69.78/68.71 kDa (mouse/rat), 69.69/72.51/69.09/59.10/64.19/64.00/85.35 kDa (human isoform 1/2/3/4/5/6/8) calculated. Human spliced isoform 3, 4, 5, and 6 are also known as isoform II, delE2/3, delE3, and delE2, respectively. Uncharacterized bands may be observed in some lysate(s).

Format: Purified

Protein G purified

Purified mouse IgG1 in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

Stable for 1 year at 2-8°C from date of receipt.

Concentration: Please refer to lot specific datasheet.

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